Supramolecular Organization Predicts Protein Nanoparticle Delivery to Neutrophils for Acute Lung Inflammation Diagnosis and Treatment

Jacob W. Myerson; Priyal N. Patel; Nahal Habibi; Landis R. Walsh; Yi-Wei Lee; David C. Luther; Laura T. Ferguson; Michael H. Zaleski; Marco E. Zamora; Oscar A. Marcos-Contreras; Patrick M. Glassman; Ian Johnston; Elizabeth D. Hood; Tea Shuvaeva; Jason V. Gregory; Raisa Y. Kiseleva; Jia Nong; Kathryn M. Rubey; Colin F. Greineder; Samir Mitragotri; George S. Worthen; Vincent M. Rotello; Joerg Lahann; Vladimir R. Muzykantov; Jacob S. Brenner

Proteins were prepared in deionized and filtered water at concentrations of 0.155 mg/mL for human albumin, 0.2 mg/mL for hen lysozyme, and 0.48 mg/mL for IgG. Crosslinked albumin nanoparticles, lysozyme-dextran nanogels, and IgG-coated liposome suspensions were prepared such that albumin, lysozyme, and IgG concentrations in the suspensions matched the concentrations of the corresponding protein solutions. Protein and nanoparticle solutions were analyzed in quartz cuvettes with 10 mm path length in an Aviv circular dichroism spectrometer. The instrument was equilibrated in nitrogen at 25°C for 30 minutes prior to use and samples were analyzed with sweeps between 185 and 285 nm in 1 nm increments. Each data point was obtained after a 0.333 s settling time, with a 2 s averaging time. CDNN 40 software deconvolved CD data (expressed in millidegrees) via neural network algorithm assessing alignment of spectra with library-determined spectra for helices, antiparallel sheets, parallel sheets, beta turns, and random coil.